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. 2012 Nov 1;139(21):4007–4019. doi: 10.1242/dev.081703

Fig. 4.

Fig. 4.

Dysregulation of cardiac genes in Irx3 and Irx5 mutant embryos. (A) Genotypes used for microarray. EGFP-positive cells were isolated by FACS. (B) Number of genes statistically (P<0.01) upregulated (orange) and downregulated (blue) in each mutant as compared with wild type. (C) Unsupervised two-way hierarchal clustering applied to the wild type, Irx3KO, Irx5KO and Irx3;Irx5 DKO samples (horizontally) and the 458 statistically differentially expressed genes (vertically). Input consists of replicate expression values for each gene and sample. Gene expression values were centered to the median. Therefore, each color patch represents fold change between expression value and the median, with a continuum from bright blue (lowest) to bright yellow (highest). Two clusters (A and B) containing genes relevant to the discrimination between Irx3;Irx5 DKO and the other genotypes are shown on the right. (D) qPCR confirmation of a number of genes identified by microarray as either downregulated, upregulated or unchanged in the Irx3;Irx5 DKO. Values are mean ± s.d. a, P<0.05; b, P<0.01; c, P<0.001; in Irx3;Irx5 DKO versus wild type. (E) The BMP pathway and genes described as upstream or downstream components of this pathway. Red arrows illustrate how these genes are misregulated in the Irx3;Irx5 DKO embryos according to the microarray data.